Methods for improving the dynamic range of biological assays

Compositions and methods are provided that extend the range of biological assays, such as immunoassays. In one embodiment a plurality of discrete test sites are used. Data for the proportion of test sites that indicate the presence of the analyte of interest and data that provides a statistical value for a signal generated by the population of discrete test sites is gathered. The results of these digital and statistical approaches are aggregated to provide an extended dynamic range. In another embodiment two or more reagent additions are provided in a serial fashion to a single test site or vessel. Such additions present different reagent sets with different dilutions of the same sample, and permit simultaneous characterization of both high and low abundance analytes in a single multiplexed assay while avoiding the high dose hook effect observed for high abundance analytes at low sample dilutions. These approaches can be combined to provide further improvements in dynamic range.